The subject matter disclosed herein relates to enhancers of amidohydrolase activity. Fatty acid amide hydrolase (FAAH) belongs to the superfamily of amidase signature proteins. FAAH enzymes are found in diverse groups of organisms including both plants and animals. FAAH enzymes hydrolyze a broad range of N-acylethanolamines (NAEs) to corresponding free fatty acid and ethanolamine, and can also act on primary acyl amides as well as acyl esters. In plants, one type of FAAH has been characterized, whereas two distinct FAAHs have been described in animal systems. The homology between plant (Arabidopsis) and mammalian (rat) FAAH proteins at the amino acid level is somewhat low over the full length of the proteins. However, the amidase signature sequence (with core catalytic residues) between plant and animal FAAH share up to 60% similarity at the amino acid level. The X-ray crystallography data of the rat protein provided new insights about the mode of action of this enzyme in NAE hydrolysis. Although no tertiary structure has been determined yet for plant FAAH, a structural homology model was developed for the amidase domain of the Arabidopsis FAAH protein and conserved catalytic residues were identified experimentally.
The activity of FAAH is a key regulatory feature of the NAE signaling pathway. The regulated accumulation of NAEs influences numerous functions in plants and animals. The functional activities and physiological effects of NAEs are mostly terminated by their degradation to FFA and ethanolamine. In plants, NAEs are involved in seedling establishment and growth. Exogenously applied N-lauroylethanolamine (NAE 12:0) arrests seedling growth and this is evident through marked changes in root architecture and elongation. This inhibitory effect of NAEs on seedling growth occurs in part through a complex interaction with ABA (abscisic acid) signaling machinery during the embryo-to-seedling transition that remains incompletely understood. On the other hand, reductions of endogenous seed NAE levels through the over-expression of FAAH in Arabidopsis results in enhanced seedling growth and increased size of roots, cotyledons and other plant organs. Other physiological processes have been attributed to FAAH mediated alteration of NAE levels in plants, such as flowering time which is induced by the expression and translocation of the FLOWERING LOCUS T (FT) protein from leaves to the vegetative meristem. FAAH over-expressing plants exhibited an early flowering phenotype in both inductive and non-inductive growth conditions, and this was associated with lower NAE levels and higher expression of FT and other key flowering genes. Still other work has attributed changes in host susceptibility to pathogens or changes in phytohormone signaling pathways with altered FAAH expression.
In animals, FAAH-mediated NAE changes are part of the so-called “endocannabinoid signaling pathway”, and this pathway plays a central regulatory role in many physiological and behavioral processes. The most widely studied NAE in animal systems is the N-arachidonylethanolamine, known also as anandamide (NAE 20:4), but other NAE species with overlapping or unique functions are known as well. As an example N-linoleoylethanolamine (NAE 18:2) or N-palmitoylethanolamine (NAE 16:0) are involved in neuron protection in the retinal ganglion cell layer against excessive extracellular glutamate and against oxidative stress for the HT22 cells, respectively. Anandamide was identified as the first endogenous ligand of the cannabinoid receptors (CB1 and CB2), and is involved in activating many of the important endocannabinoid pathways. Anandamide and other NAEs have been associated with different processes such as pain modulation, memory, anxiety, appetite, etc. Their levels are controlled largely through hydrolysis by FAAH. Thus, FAAH has become a major therapeutic target for many disorders that involves NAE signaling in situ.
Several approaches have been employed to increase the level of NAEs in plants or animals including the direct application of NAEs or pharmacological reagents that inhibit NAE degradation. Utilization of general and/or specific inhibitors of FAAH activity such as phenylmethylsulfonyl fluoride (PMSF), 5Z,8Z,11Z,14Z-eicosatetraenyl-methyl ester phosphonofluoridic acid (MAFP) or 3′-(aminocarbonyl)[1,1′-biphenyl]-3-yl)-cyclohexylcarbamate (URB597) have been reported to elevate endogenous levels of NAE, and to extend or amplify processes regulated by NAE signaling. Genetic approaches have been developed to reduce FAAH expression (FAAH knockouts) in mice and Arabidopsis, although this approach has shown limited success, especially in plants where it appears that there are redundant pathways for NAE catabolism, and where it has been difficult to raise endogenous NAE levels dramatically in vivo. On the other hand, it has been possible to over-express FAAH in plants and reduce NAE levels to some extent to influence several physiological processes including growth, defense, and flowering. However, there is limited information on chemical compounds that reduce the NAE content in plant and animal systems via enhanced FAAH activity.
Among the multitude of renewable resources, cashew nutshell liquid (CNSL) is an important by-product of the cashew nut industry that is currently used for green chemicals and technologies. More than 32% of the cashew shell is CNSL, the key constituent of CNSL being cardanol, a bio based non isoprene lipid, comprising a rich mixture of phenolic lipids: 5% of 3-(pentadecyl)-phenol (3-PDP), 50% of 3-(8Z-pentadecenyl)phenol, 16% of 3-(8Z,11Z-pentadecadienyl)phenol and 29% of 3-(8Z,11Z,14-pentadecatrienyl)phenol. Cardanol's unique properties stem from the varying degree of cis-double bonds and an odd number of hydrocarbons with easily accessible saturated and unsaturated hydrocarbon chains.
The discussion above is merely provided for general background information and is not intended to be used as an aid in determining the scope of the claimed subject matter.